Download ETP: The Immune System

ETP: The Immune System
Ellen Fasman
IISME: Summer 2008
Main Teacher Objective: to enhance student understanding of immunology,
as well as interest
powerpoint.
Students will do a variety of labs, activities, and present a
Specific objectives: to explain the structures and functions of the immune system. This
includes concepts such as hematopoiesis, innate and adaptive immunity, the
inflammatory response, antigen-antibody reaction, and vaccines. Labs will include
microscopy using prepared slides of blood smears and pathogens, the ELISA test, blood
typing, making serial dilutions, and PCR. Labs will highlight the importance of
replicates and controls in an experiment
Grade levels: 9th and 10th grade Biology, and
grade 10-12 Physiology students .
Standards covered:
My ETP will focus on Standard #10 of the California State Standards. This includes the
following:
10a. students know the role of skin in providing nonspecific defenses against infection.
10b. Students know the role of antibodies in the body's response to infection
10c. Students know how vaccination protects an individual from infectious diseases.
10d. Students know there are important differences between bacteria and viruses with
respect to their requirements for growth and replication, the body's primary defenses
against bacterial and viral infections, and effective treatments of these infections.
10e. Students know why an individual with a compromised immune system (a person
with AIDS) may be unable to fight off and survive infections by microorganisms that are
usually benign.
10f. Students know the roles of phagocytes, B-lymphocytes, and T-lymphocytes in the
immune system
Materials needed: micropipettes with tips, centrifuge, ELISA kits with reagents,
antibodies, incubator, freezer, ABO Rh kits
Lectures and labs will go together
Week 1: Innate and Adaptive Immune Systems
Week 2: Structures and functions of the immune sytem, hematopoiesis
Week 3: Blood typing and ELISA (antigen/antibody reactions)
Week 4: Student projects/assessments
3 PPTs
1. The Immune System
2. The Inflammatory Response
3. Vaccines
Topics Covered in PPTs
a. Describe the organization of the Immune System and Lymphatic System
b. Describe the protective functions of the skin and mucous membranes
c. Explain hematopoiesis
d. Explain the importance of phagocytes and natural killer cells
e. Compare body defenses: non-specific (innate) vs. specific (adaptive)
immune systems
f. Describe the inflammatory response
g. Describe ways in which antibodies act against antigens
h. Compare and contrast the development of B and T cells
i. State the roles of B and T cells
j. Explain how antigen-presenting cells (macrophages) function
k. Explain T cell activation and interactions with other cells of the immune
response
l. Describe how vaccination works
m. Explain how the ELISA test works, in addition to its uses, and limitations
4. Assigned Readings:
a. Textbook: Biology, Prentice-Hall Publishing Company, 2007.
b. Textbook: Essentials of Human Anatomy and Physiology, Marieb, Elaine,
Pearson Education, Inc, Eighth Edition, 2006.
5. Activities
a. Immune cell posters: B cells, Helper T cells, monocytes, phagocytes,
Natural Killer cells, antigen presenting cells, plasma cells(plus antibody
group); Groups of 4; jigsaw the parts
b. Student PPTs on immune system topics (possible topics: Asthma,
Hemolytic Disease of the Newborn, Anaphylaxis, Lupus, Eczema,
Crohn’s Disease, allergy, allergy shots, influenza vaccine, smallpox
vaccine, poison ivy, HCG pregnancy test, HIV antibody test)
6. Labs:
a. ABO/Rh blood typing
b. Microscopy: look at stained cells, hematocrit
c. Micropipetting and making serial dilutions
d. Bodily Fluids Lab (ELISA test)
Name:
Date:
Period:
The Immune System
Pre-Test
1. What is the body’s first line of defense?
2. What are pathogens? Give three examples.
3. What are the names of the cells of the immune system?
4. In what organs are the cells of the immune system made?
5. What is innate immunity? Give an example.
6. What is adaptive immunity? Give an example.
7. Define antigen.
8. How do antibodies help to defend the body?
9. Differentiate between cellular and humoral immunity.
10. What is a vaccine made of? How does it work?
Bonus Questions:
Mr. Brown, an 80-year old man, is complaining about having to receive a flu shot every
year. Why is it necessary to receive these shots every year?
Name:
Date:
Period:
The Immune System
Post-Test
11. What is the body’s first line of defense?
12. What are pathogens? Give three examples.
13. What are the names of the cells of the immune system?
14. In what organs are the cells of the immune system made?
15. What is innate immunity? Give an example.
16. What is adaptive immunity? Give an example.
17. Define antigen.
18. How do antibodies help to defend the body?
19. Differentiate between cellular and humoral immunity.
20. What is a vaccine made of? How does it work?
Bonus Questions:
Mr. Brown, an 80-year old man, is complaining about having to receive a flu shot every
year. Why is it necessary to receive these shots every year?
Bodily Fluids Lab
Objectives
To learn:
* how to perform an ELISA assay
* the use of antibodies in research and in diagnostic/forensic tests
* how an infectious agent is spread through sharing of “bodily fluids”
* the importance of replicates and controls in an experiment
Summary
Students use an ELISA assay to determine whether they are carriers of an
“infectious agent.” The lab is a simulation; no infectious agents or bodily fluids
are used.
Background Information
The enzyme-linked immunosorbent assay, ELISA, is a test used to detect and
quantify specific antigens in a mixture; this mixture could be blood, urine, semen,
or other bodily fluids. It is the basis for many diagnostic tests, including the home
pregnancy test and the HIV test for AIDS. In the ELISA test, protein (either
antigen or antibody) is bound in wells of an assay plate. The assay plate is made
of polypropylene and is specifically made for binding to proteins. We will use the
antibody-capture assay method.
The steps of the general ELISA test are:
1. The sample (bodily fluid) that may contain the antigen of interest is added to
a polypropylene plate with wells.
2. The excess sample is removed and wells are washed.
3. Add enzyme-tagged antibodies, which are directed against the antigen of
interest.
4. Wash the unbound antibodies.
5. Add a color substrate.
6. View the result: if there is a color change, then the sample contains the
infectious agent (antigen of interest).
Experimental Procedure
Introduction:
This experiment contains two parts. In the first part, each student will share his or her
“bodily fluids” with three other students. Then each student will perform an ELISA
assay on the shared fluids. Each group will run positive and negative controls on a group
plate. After recording the data, each student will determine if he or she is infected and
the total number of students infected with the “disease agent.”
Kit Materials:
Solutions:
20X PBS
10% Tween 20
10X Sodium Carbonate Solution
Antigen (BSA $ biotinylated-BSA)
1X PBS with BSA
Streptavidin-peroxidase
1X Citric Phosphate Buffer
TMB
50 mL
10 mL
20 mL
150 mL
10 mL
2 uL
20 mL
tablet
Abbreviations used in this lab:
PBS
BSA
mL
ul
mg
min
TMB
H2O2
Ag
Ab
Tween 20
Phosphate-buffered saline
Bovine serum albumin
milliliter
microliter
milligram
minute(s)
3,3’,5,5’-tetramethylbenzidine
hydrogen peroxide
antigen
antibody
polyoxyethylenesorbitan monolaurate
Part 1: Preparing the Samples
Materials:
Clear “bodily fluids” tubes
Yellow microcentrifuge tube (for unshared sample)
Blue microcentrifuge tube (for shared sample)
Transfer pipette
Marker
Rack for microcentrifuge tubes
Procedure
1. Label a yellow tube and a blue tube with your initials. Divide the “bodily fluids”
sample (clear tube) evenly between the yellow and blue tubes using a transfer
pipette.
2. Place the yellow tube into the rack at your lab station. This is your unshared
sample.
3. Share your sample in the blue tube with another student. To do this, use a transfer
pipette to combine the two samples into a single blue tube. Then divide the
sample evenly between the two blue tubes. Throw out the transfer pipette.
Record the name of the person with whom you share fluids on the form below.
4. Find a second person with whom to share fluids and repeat step #3. Again, record
the name and throw the transfer pipette away.
5. Find a third person with whom to share and repeat step #3. Your blue tubes should
now contain the “bodily fluids” from 4 different people. You should have 3
names recorded on your form.
Record of Sample Sharing
Your name: ________________________________
Students with whom you shared “bodily fluids:”
1.______________________________________
2.______________________________________
3.______________________________________
Part 2: ELISA Assay on Shared Samples
In this part of the lab, you will run an ELISA test on your shared samples. Each sample
will be in triplicate and you will share an assay plate with your lab partners.
Materials
Quantity
Mixed “bodily fluids” in blue tube
ELISA plate
1 tube per student
1 plate per group
Green tube (positive control sample)
Pink tube (negative control sample)
Orange tube (antibody solution)
Purple tube ( color substrate)
Squeeze bottle with wash buffer
Stack of paper towels
50 uL fixed volume pipette
Yellow pipette tips
Marking pen
Rack for microcentrifuge tubes
1 tube per group
1 tube per group
1 per group
1 per group
1 bottle per group
1 stack per group
1 per group
10 per group
1 per group
1 per student
Procedure:
1.
Using the 50-uL fixed volume pipette with a new tip, each person in the group
adds 50 uL of shared “body fluid” (blue tube) into each of 3 wells. Record the
location of each person’s wells on the ELISA assay form.
2.
Using a fresh pipette tip for each reagent, prepare 3 positive (green tube) and 3
negative (pink tube) control wells on your plate. Record the location of the
controls on the ELISA assay form. Note: the control wells should be adjacent to
the experimental wells—not separated by empty wells.
3.
Let the plate sit for 15 minutes at room temperature. While the plates are
incubating, write your name in the table on the board, as instructed by your teacher.
4.
Shake off the fluid into the sink. To empty the wells completely, flick the plates
(well side down) against the edge of the sink.
5.
Use the squeeze bottle of the wash solution to wash the wells 3 times, shaking out
the wash buffer between washes.
6. Tap the plate upside down firmly no a stack of paper towels to remove excess fluid.
7.
Using a new pipette tip, add 50 uL of antibody solution (orange tube) to each well.
Let the plate sit for 5 minutes at room temperature.
8.
Shake off fluid and wash the plate as in steps 4-6 above.
9.
Last step: Using a new pipette tip, add 50 uL of color reagent (purple tube) to each
well. Let the plate sit for 10 minutes at room temperature.
10. Observe the results. Negative wells (“uninfected” and the negative controls) will
remain totally clear. Positive wells (“infected” and the positive controls) will turn
blue.
11. Go to the table where you wrote your name and place a plus or a minus next to
your name, depending on your results. Also, put a plus or a minus next to the
names of the 3 people with whom you shared “bodily fluids.” For example, if
your result was positive, you mark a plus sign (+) next to your name and 3 other
names and if your result was negative, you mark a negative sign (-) next to your
name and 3 other names.
Sample Loaded
(Student Name)
Results
(infected/non-infected)
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
____________________
Positive controls
___________________
____________________
Negative controls
___________________
____________________
Triplicate Assay
Mark (+) or (-)
Simple Procedure (at lab bench)
1. Each student receives a clear tube of “bodily fluid.” Put aside half the sample
for later testing into a yellow tube.
2. Share your “bodily fluid” with another student by combining them in a single
tube, mixing, and splitting them back into 2 tubes again.
3. Now share your “bodily fluid” with 2 more students in the same way.
4. Pour shared sample into a blue tube for testing.
5. To confirm results of the first experiment, the unshared samples can be
assayed.
Questions
1. What is the full name of the test we simulated in the laboratory?
2. What is the purpose of this experiment?
3. What antibody “tag” does an ELISA use?
4. What’s the difference between a positive and negative test?
5. Why did we run positive and negative controls with the assays? Why did we run
them on every plate, instead of just one?
6. What is meant by a “false positive” result? What is one way you could get a false
positive result?
7. What is meant by a “a false negative” result?” What is one way you could get a
false negative result?
8. Why did we assay each sample in triplicate?
9. Did your 3 wells for each sample have exactly the same intensity of blue color?
10. Why did you wash the plate before you added a new reagent? What could happen
if you didn’t wash the plate?
11. How many students in the class were “infected” by the disease agent?
12. How could you figure out who the original “infected” person is?
13. What do your results say about the spread of disease through activities in which
bodily fluids are shared?
14. After doing the lab, would you agree or disagree with the following statement:
“When you have sex with someone, you are also having sex with everyone that
they have previously had sex with.” Explain your answer.
Activities
1. Immune cell posters:
B cells, T cells, monocytes, phagocytes, Natural Killer cells, antigen presenting
cells, Cytotoxic T cells; Groups of 4; jigsaw the parts
Poster Requirements:
1. Draw cell and label nucleus, cell membrane, 3 organelles
2. Show the cell in action; it must be interacting with something else (foreign
antigen, pathogen, cell, etc).
3. Explain, in steps, the cell in action. What is it reacting to? Specify what it’s
doing and if this is an example of innate or adaptive immunity.
4. How is this cell different from all other cells? How is it similar?
2. Student PPTs on immune system topics
(Possible topics: Asthma, Hemolytic Disease of the Newborn, Anaphylaxis,
Lupus, Eczema, Crohn’s Disease, allergy, allergy shots, influenza vaccine,
smallpox vaccine, poison ivy, HCG pregnancy test, HIV antibody test)
PPT Requirements:
Disease
1. Summary statement: how is this disease significant in the field of immunology?
2. What is the cause of the disease?
3. How does the disease work (symptoms)?
4. Which cells and antibodies of the immune system are involved in the disease?
5. Treatment of the disease
Pregnancy, HIV Test:
1. How is this test significant in the field of immunology?
2. What is the reason for the test? When and to whom is it given?
3. How does the test work?
4. Which cells and/or antibodies are involved?
5. What are the limitations and dangers of the test?
-Advertisement-
Serial Dilutions Made Easy
Author: Jan Hilten and Carol Sanders
Woodrow Wilson Biology Institute
1993
Introduction:
Many areas of science use serial dilutions in the preparations for different experiments.
This exercise is presented as an aid for the teacher in helping his/her students improve
their skills and more quickly understand the particular application for which serial
dilutions are a tool. Serial dilutions are often used in microbiology, biotechnology, and in
chemistry classes, to name just a few. Therefore a clear, concise, and non-threatening
approach to the learning of this very important concept is essential.
Serial dilutions are usually made in increments of 1000, 100 or 10. The concentration of
the original solution and the desired concentration will determine how great the dilutions
need to be and how many dilutions are required. Important also is the total volume of
solution needed. If only small quantities of solutions are needed then greater numbers of
dilutions are necessary.
The most common examples deal with concentration of cells or organisms, or the
concentration of a solute. The approximate concentration should be known at the start of
the experiment before the appropriate number and amount of dilutions can be made. In
order to arrive at the desired concentration, use serial dilutions, instead of making one big
dilution, in order to finally arrive at the desired concentration. This method is not only
cost effective but it also allows for small aliquots to be diluted instead of unnecessarily
large quantities of materials.
This technique involves the removal of a small amount of an original solution to another
container which is then brought up to the original volume using the required buffer or
water. In the example below, if you have 1 mL of your original solution, and you remove
10 µL and place it in a tube containing 990 µL of water or media you have made a 1:100
dilution. If the original solution contained 5 x 108 organisms or cells/mL, we now have a
concentration of 5 x 106 cells/mL, because we have simply divided our concentration by
100. Now, if we want to dilute this by a factor of 1:1000, we must remove 1 µL of the
second solution and place it in a tube containing 999 µL of media. We have now diluted
our secondary concentration by 1000, and would then divide our concentration by 1000 to
yield a 5 x 103 cells/mL.
Practice Exercises
1. You are given a test tube containing 10 mL of a solution with 8.4 x 107 cells/mL.
You are to produce a solution that contains less than 100 cells/mL. What dilutions
must you perform in order to arrive at the desired result?
ANSWER: You should perform a series of three 1:100 dilutions to yield 84
cells/mL.
1 mL of original solution to 99 mL of water = 8.4 x 105 cells/mL.
1 mL of second solution to 99 mL of water = 8.4 x 103 cells/mL.
1 mL of third solution to 99 mL of water = 8.4 x 101 or 84 cells/mL.
2. You have a microtube containing 1 mL of a solution with 4.3 x 104 cells/mL and
you are to produce a solution that contains 43 cells/mL. What dilutions must you
perform?
ANSWER: You could perform the following dilutions:
10 µL of original solution to 990 µL of water = 4.3 x 102 cells/mL.
100 µL of second solution to 900 µL of water = 4.3 x 101 or 43 cells/mL.
3. You are given a container with 5 mL of a solution containing 5.1 x 103 cells/mL.
You are to produce a solution that contains approximately 100 cells/mL.
ANSWER: You would perform the following dilutions:
0.5 mL of original solution to 4.5 mL of water = 5.1 x 102 cells/mL
1 mL of second solution to 4 mL of water = 1.02 x 102 cells/mL or 102 cells/mL
4. You are given a container of yeast cells for which Klett units have been
determined on a Klett Summerson Colorimeter. The container contains a
population whose concentration is 2.6 x106 cells/mL. You are to prepare a
suspension which, when you spread 1 mL of the suspension on appropriate media,
will result in about 100 cells.
ANSWER:
10 µl of original solution to 990 µl (or 1.0 mL) of sterile water = 2.6 x 104
cells/mL
10 µl of second solution to 990 µl (or 1.0 mL) of sterile water = 2.6 x 102
cells/mL
0.5 mL of third solution to 0.5 mL of sterile water = 1.3 x 102 or 130 cells/mL
0.77 mL of fourth solution to 0.23 mL of sterile water = 100 cells/mL
TITLE OF VIDEO:
An Inside Look: The Flu
VIDEO COMPREHENSION QUESTIONS:
1. What happens immediately after viruses and bacteria enter the body?
2. How does a virus take advantage of the way human cells work?
3. Why does the entire body rather than just the throat experience flu symptoms?
4. How does a T cell become activated, and what happens immediately after it's been
called into duty?
5. What are antibodies, and what is their role in fighting the flu?
6. What causes immunity to the flu?
An Inside Look: The Flu
VIDEO COMPREHENSION QUESTIONS AND ANSWERS:
1. What happens immediately after viruses and bacteria enter the body?
When viruses and bacteria first enter the body through the nose, they encounter the hairs
lining the nostrils. These hairs trap nearly every particle that's inhaled, and they contain
mucus that dissolves bacteria. A virus may survive, however, if it becomes detached from
a nose hair and is sucked deeper into the nose. From the back of the nose, the virus can
either go to the stomach, where it will be destroyed, or make its way to the throat,
beginning the process of giving the person the flu.
2. How does a virus take advantage of the way human cells work?
It impersonates one of the proteins that cells use to communicate with each other. A cell
can therefore be fooled into thinking the virus is a harmless protein and will permit the
virus to enter it. Once inside the cell, the virus begins to use the cell's structure to
manufacture components for new viruses.
3. Why does the entire body rather than just the throat experience flu symptoms?
As the flu viruses multiply, macrophages in the throat release interleukins, which are
chemicals that go through the bloodstream to summon reinforcements to the throat. The
interleukins cause nerves to be hypersensitive, making even slight touches uncomfortable.
They also raise the body's temperature to make it a less hospitable environment for the
virus. This temperature increase tricks the body into feeling cold. Blood vessels around
the brain swell, causing a headache. All of these symptoms help remind the flu victim to
slow down so that energy can be channeled into defeating the virus.
4. How does a T cell become activated, and what happens immediately after it's been
called into duty?
Dendritic cells gather fragments of the flu virus and then search the body's lymph glands
for an appropriate T cell. Once this T cell has been located, it begins to divide. The new T
cells then move to the throat through the bloodstream and selectively destroy the infected
throat cells.
5. What are antibodies, and what is their role in fighting the flu?
Antibodies are tiny proteins manufactured by B cells. They lock onto the spikes of newly
produced viruses, paralyzing the viruses and preventing them from infecting new cells.
6. What causes immunity to the flu?
Some T cells remain as memory cells, patrolling the body for the rest of the person's life.
If the virus tries to enter the body again, these memory cells will instantly wipe it out,
unless it has mutated.
Pathogen Project
Objectives: to learn about a disease: cause, symptoms, diagnosis, prognosis, mode
of transmission, epidemiology, and vaccines (if any)
Instructions:
Use a variety of resources, including books, periodicals, the Internet, health care
professionals, and the local department of public health
Below is a list of suggested diseases to research:
Polio
Pneumonia
Botulism
Salmonella
Meningitis
Herpes 1
Herpes 11
Scarlet Fever
Tuberculosis
Staphylococcus aureus
Syphilis
Mumps
Lyme Disease
Encaphalitis
Pseudomonas
Pyelonephritis
Chicken Pox
Small Pox
Rubella
Yellow Fever
Rabies
Mononucleosis
Tetanus
Gonorrhea
Lupus
Anthrax
Diphtheria
Ebola
Streptococcus pyrogenes
Format:
Poster and speech or powerpoint (including oral presentation)
Parts of the Project:
1. Summary Statement: name of disease, standout features, part of the body which is
affected
2. Organism causing disease: virus, bacteria, parasite, fungus? Give the genus and
species or viral name
3. Symptoms: List the characteristic signs of the disease. Do new symptoms develop
as the disease progresses? What happens if the disease goes untreated?
4. Diagnosis: what tests are performed by doctors to diagnose the disease?
5. Mode of transmission: How is this disease passed from one person to another?
6. Treatment: Consider all aspects of treatment, including medication, physical
therapy, rest, etc. Mention any treatments which are not based in Western
medical tradition; is there evidence for the effectiveness of these treatments?
Why do you think they may work?
7. Vaccines: Are they available for the disease? If yes, what type of vaccine is it?
What is the recommended schedule for vaccination to prevent this disease? Does
the vaccine have any side effects?
8. Epidemiology: Who gets the disease? Mention distribution of disease, based on
factors such as age, sex, race, religion, lifestyle, and geographical area.
9. Prognosis: What is the chance of being cured if you contract this disease? If
cured, could you get the disease again?
Medical Detective
A Walk in the Woods
Jack is a lawyer who lives in a wooded suburb near Stony Brook, New York He
has two children, the youngest being 25 years old. He typically works 50 hours a week.
In his spare time he likes to go fishing. His house, a 100 acre estate, is tucked away
from the noise and commerce of the city. His daily routine includes going for a walk
through the woods and fields of his property with his Golden Retriever, Lancelot.
Jack is also an avid birder and usually brings his binoculars with him. .
Jack grew up in inner city Pittsburgh in an area where there were a lot of
factories and textile mills. Jack had always been healthy but one of his two siblings
died of leukemia. The other was chronically ill and the doctors could never come up
with a specific diagnosis.
In early August, Jack began to feel run down. He noticed that he had been
having more headaches than usual. He didn’t go to work for three days due to a fever
and general malaise. He felt better for a few days and then missed another two days of
work with a low grade fever, night sweats and chills, and severe back pain.
Back at work, Jack’s coworkers worried about him. They noticed that he
appeared sluggish and without his usual high spirits. They also noted that he was more
apt to forget things and had a hard time focusing on important matters at work.
Jack quit working because he had worsening pain in his neck and shoulders and
he felt too exhausted. He felt a strange weakness in the left side of his body. What was
once a small pink dimple on his right leg now looked like a round red rash with a
smaller rash in the center. He became depressed and began staying in bed longer and
longer periods. Finally, his wife convinced him to see a doctor.
Now you be the doctor. What do you think is wrong with Jack?
Teacher’s Guide: A Walk in the Woods
Disease: Lyme Disease
Relevant facts:
Lives in wooded area- where there are deer, ticks, mosquitoes
Location: NY: site of an epidemic of Lyme Disease
Time of year: August: breeding time for ticks
Symptoms: malaise, fever, muscle weakness, lack of sensation on one side,
depression, outwardly expanding skin rash/bullseye rash (erythema migrans)
Medical Detective #2
Kids on Fire
Joey is a 5-year old boy who lives with his two brothers in a suburb of
Minneapolis. He was born with deafness in his right ear. Joey started kindergarten in the
Fall and goes to the school day care center from 12:00 PM until 5:00 PM most school
days. It’s a fairly crowded day care, 20 kids on average. Three kids were reported sick
this week, all with high fevers.
Two days a week, Joey plays soccer at the local elementary school. He had a
mild concussion two months prior after heading a ball in a championship game. Joey’s
parents both work full-time, and they just had a new swimming pool installed in their
backyard, equipped with a hot tub. After a birthday party for his oldest party at their pool,
Joey suddenly felt hot. In fact, that night his fever jumped to 103 degrees. After some
Tylenol and a lukewarm bath, it went down to 101. However, his mother was still
worried. The next morning Joey’s fever spiked again, this time to 105 degrees. He
complained of a stiff neck and said there were snakes crawling all over him. Joey’s
mother rushed him to the hospital. While he sat in the waiting room, the bright lights
made him scream. He said his head felt like it was exploding
Sadly enough, a secretary from Joey’s school called to tell her that one of the
boys at Joey’s day care suddenly died. Five years later, Joey has remnants of the disease:
weakness on one side of his body and loss of the tips of two toes.
Now you be the doctor. What do you think is wrong with Joey?
Teacher’s Guide: Kids on Fire
Disease: Bacterial Meningitis
Relevant clues:
Goes to daycare with lots of kids, where infections spread easily
Two kids reportedly with high fevers, one died
Symptoms: high fever, stiff neck, hallucinations (snakes crawling over him), light
sensitivity
Lasting effects: loss of the tips of his toes/ weakness on one side of his body
Medical Detective
A Boy Named Chile
Chile Juarez is an eighth grader living in Sylvester, Georgia. He is the youngest
of three brothers and lives with his mother and grandmother in an apartment above a
pawn shop. He likes to play baseball or skateboard at the park with kids who live
nearby. He has a stray cat, named Reggie, who came into his life when he was nine
years old. Reggie is an outdoor cat who seems to have been in a lot of fights, owning to
his numerous scars and “foxhole air.”
Chile is an excellent student and his mother dreams of being able to afford
college for him someday. She figures by the year 2011, she’ll be able to work fulltime and won’t have her elderly mother to take care of. She works at a local factory
which makes truck parts. Chile’s father left when he was only one.
When Chile was ten, he came home from school complaining of stomach cramps.
After dinner, he experienced more cramps and diarrhea. For the next two days, he
lay in bed, and couldn’t hold down any food. His mother did not have medical
insurance, so she treated Chile with the best she knew how: cool compresses,
Tylenol, and, of course his favorite snack of peanut butter and honey sandwiches.
After about one week, his symptoms subsided and he felt his old self again.
Four months later, Chile began having terrible pains in his wrists and heels. He
stopped urinating. This went on for seven days. Chile’s mother went to Wal-Mart
and picked up some aspirin to relieve the pain. His mother brought him to the
emergency room after noticing the whites of his eyes turned yellow. At the
hospital, his fever spiked to 103.7; Chile’s mother told the doctor about his
previous bout with upset stomach and diarrhea. They decided to do a blood and
urine test, including a HLA-B27 ELISA, plus a spinal tap. The doctors
concluded that he was suffering from complications of his untreated illness six
months prior.
Now.. you be the doctor…What disease did Chile have?
Teacher’s Guide: A Boy Named Chile
Disease: Salmonella Poisoning
Symptoms: stomach cramps, diarrhea, fever, joint pain,
Later symptoms: yellow white of eyes (jaundice), lack of urination (kidney failure)
are later effects of untreated Salmonella poisoning—all part of Reiter’s Syndrome
Other clues: mother gives him peanut butter sandwiches; there was an outbreak of
salmonella contamination in Peter Pan peanut butter from Wal-Mart stores in
Sylvester, Georgia in 2007
HLA-B27 ELISA test done- the major test for salmonella poisoning
Medical Detective:
Bea Stung
Bea Hopstone lives in the outskirts of Central Park in New York. She is a
freelance writer for the New York Times and the New York Herald Tribune and
co-author of History of Dictatorship. She travels frequently to Thailand, where
she covers the recent political tension on the Thai-Cambodian border. Born and
educated in England, she prefers the rattle and race of New York City to her quiet
life in Chelsea. She is 36, a mother of three children, twin girls, aged 4 and a boy,
aged 8. Her husband is an architect. Bea enjoys big band jazz, tennis, and long
walks with her family through Central Park. She is an excellent ping pong player
and regularly plays in tournaments on Canal Street with a team called the Barons
of Bounce. She has tendonitis in her right elbow and asthma.
Bea was working on an article one sunny September morning in her
uptown office, eating a corned beef sandwich with one hand, typing in the other.
Over the course of the afternoon, she experienced a headache which wouldn’t
stop and nausea. She went home early and lay in bed for what seemed like a
couple of hours (It was actually 12). Her family assumed she was exhausted, since
she was working more hours than usual, covering a feature story on the recent
spike in death rate among crows and blue jays in New York City. Her headache,
now much worse, had spread to her back, and her limbs felt especially weak. Her
husband took her temperature: 39.4 C. “My head feels like it’s on fire!,” she
yelled. He rushed her to the hospital.
The doctor, a specialist in tropical diseases, noted the following symptoms:
fever, frontal headache, diaphoresis, lymphadenopathy, severe vomiting and
diarrhea, bilateral leg weakness, neck stiffness, and severe eye pain. Over the
next six days, a broad spectrum antibiotic was administered, which didn’t relieve
the most severe symptoms of the disease. A morphine drip through an I.V. was
given for pain relief. Below is a summary of the test results for Bea Hopstone:
Vital Signs:
HEENT: photophobic
Blood Pressure: 126/80
Neck: nuchal rigidity
Pulse: 96
Respirations: 18
Temperature: 39.4 C, oral
Neurological Exam: demonstrates an alert and
oriented middle-aged female with normal cranial
nerves 1-X11
Test
Date
Lumbar Puncture
09/04/1999
Results
Clear CSF fluid, normal glucose,
mildly elevated protein
Absence of pathogens using gram stain
and culture
CBC
09/04/1999
CSF lymphocyte count: 158 cells/uL
Total WBC: 10,000 cells/uL
Lymphocyte: 4,000/uL
Hemoglobin: 11.9g/dL
Hematocrit: 33.1%
Normal
Cranial CT scan
09/04/1999
CSF IgM ELISA
09/10/1999
Toxicology Screen (urine)
09/04/1999
Positive for something else
No alcohol or illicit drug use
Chest X-ray
09/10/1999
Minimal hyperventilation
MRI
09/10/1999
Leptomeningeal enhancement of the
conus medullaris and cauda equina
Negative for Herpes, AIDS,
cytomegalovirus
Two days later, Bea became less responsive and her respiratory muscles seemed to be
working overtime. She stopped talking and had to be put on a respirator.
Now you be the doctor. What do you think is wrong with Bea?
Teacher’s Guide:
Bea Stung
Disease: Viral Encephalitis caused by West Nile Virus
Relevant data:
increase in mortality rate for blue jays and crows- vectors of West Nile Virus-90%
crows killed in 1999 from West Nile Virus
New York City- site of West Nile Virus epidemic
Bea likes to walk in the park (near mosquitoes, birds)
WNV is taken in by mosquitoes, birds; Bea was most likely bitten by an infected mosquito
Symptoms: fatigue, fever, joint pain, loss of memory and consciousness, eye
pain, high lymphocyte count, clear CSF (It’s milky in bacterial infections
Vital Signs:
Signs of WNV encephalitis:
High pulse
High body temperature
Nuchal rigidity
Photophobia
Other clues of WNV viral encephalitis:
Antibiotics didn’t work- not bacterial
Gram stain and culture-negative--- not bacterial
Lab tests:
Clear CSF: indication of non-bacterial infection
High CSF lymphocyte count (There should be none)- indication of infection
CBC: Low RBC, high lymphocyte count
MRI shows upper and lower spinal cord swelling
ELISA test positive for something else: viral encephalitis
Case Study Directions
1.
2.
3.
4.
5.
6.
Read the case study a couple of times. Underline what you think are relevant
pieces of information. NOT EVERYTHING IS RELEVANT.
Make a list of the relevant facts on the Case Study Notes sheet.
Research the symptoms.
Make a list of “Rule outs” and “Possibilities.” Explain why you rule it out or
consider it a possibility.
Write a one paragraph essay on which disease you think the patient has. Explain
which symptoms support your hypothesis and which symptoms are absent,
according to the literature.
IT’S OK IF YOUR DISEASE IS INCORRECT. YOU WILL STILL GET
CREDIT IF YOUR REASONING IS LOGICAL AND WELL-RESEARCHED.
Case Study Notes
Relevant Facts/
Patient Background
(age, sex, health, lifestyle,
place of residence,
occupation, etc)
Bibliography
Symptoms
Research on symptoms
Rule Outs
Possibilities
Model of the Immune System Project
Objective: to make an interactive model of either Humoral Immunity or Cell-mediated
Immunity and LEARN what’s going on
Materials
use any objects to represent parts of the immune system
Parts of the Project:
Model:
1.
Create a “working” (interactive) model of either Humoral or Cell-mediated
immunity. Each member of the group should be able to explain each structure
and homeostatic function.
Write-up:
2.
3.
On a sheet of paper, draw the model. Label each structure and process according
to its scientific name.
Write a one paragraph summary of how your model works. How is it significant
to biology?
Directions for Model:
1. Make a moveable model showing the “cascade effect” in either humoral or cellmediated immunity. That is, show how one thing affects another and another
down the line. A “domino effect” is one way of thinking about the adaptive
immune response.
2. The items in boxes represents different cells. The items on arrows are actions.
3. The structures and actions should be as realistic as possible.
Humoral Immunity
T Helper Cell
-
activates
B
Cell
Plasma
cell
differentiates into
Becomes
Bacterial
cell
stimulates
differentiates into
infects
Memory
Cell
releases antibodies
and binds to
engulfs and digests
Macrophage
Bacteria covered
with Antigen
Cell-Mediated Immunity
releases cytokines
Antigen-Presenting Cell
(APC)
Cytotoxic
T Cell
T Helper Cell
activates
and activates
differentiates
infects APC cell
Bacterial
cell
into
Infected host
cell
phagocytosis
macrophage
T Memory CellMigrates to lymph nodes to be
activated quickly during a
second invasion
References:
http://www.accessexcellence.org
http://www.biology.arizona.edu
Kristi DeCourcy, The ELISA Assay: An Immunological Experiment, Information
Manual, Fralin Biotechnology Center, Virginia Tech, 2003
http://www.discoveryschool.com
Harlow,E., and Lane,D.1988. Antibodies, a Laboratory Manual. Cold Spring Harbor
Laboratory, Cold Spring Harbor.
http://infectious diseases.about.com
http://www.informaworld.com, “Biomarkers,” 2007
Elaine M Marieb, Essentials of Human Anatomy and Physiology, Pearson Benjamin
Cummings, 2006
Peter Parham, The Immune System, Garland Science Publishing, 2005
www.RnDSystems.com, Human CCL17/TARC, DuoSet ELISA Development System,
R&D Systems Europe,Ltd.
Acknowledgements:
Roche Palo Alto, Mentors: Jennifa Gosling and Mirella Lazarov
Roche Palo Alto Staff: Dave Morris PDEI, Dan Cooper PDEI, Rudyard Ress PDEI,
Summer IISME Internship, 2008